Department of Neurology, University of Michigan and VA Ann Arbor Healthcare System, Ann Arbor, MI 48109, USA.
Gene Ther. 2012 Nov;19(11):1107-13. doi: 10.1038/gt.2011.188. Epub 2011 Nov 17.
We previously reported regulated expression of erythropoietin (EPO) over 4 weeks in the peripheral nerve in vivo, using a herpes simplex virus (HSV)-based vector containing a Tet-on regulatable gene expression cassette. To create a vector that would be appropriate for the treatment of chronic neuropathy, we constructed a HSV vector with expression of EPO under the control of the Tet-on system in which the HSV latency-associated promoter 2 element was used to drive the expression of the Tet-on transactivator. EPO expression from the vector was tightly controlled by administration of doxycycline (DOX) in vitro. One month after inoculation of the vector to transduce dorsal root ganglion (DRG) in vivo, administration of DOX-containing chow-induced expression of EPO. Mice with streptozotocin-induced diabetes, inoculated with the vector, were protected against the development of neuropathy by continuous administration of DOX-containing chow over the course of 3 months. Identical results were achieved when DOX was administered every other week over 3 months of diabetes, but administration of DOX, 1 week out of 3, provided only partial protection against the development of neuropathy. Taken together, these results suggest such a vector is well suited for clinical trial for the treatment of chronic or subacutely developing neuropathy.
我们之前曾报道过,使用含有 Tet-on 可调控基因表达盒的单纯疱疹病毒(HSV)载体,可在体内外周神经中实现长达 4 周的红细胞生成素(EPO)调控表达。为了构建适用于治疗慢性神经病变的载体,我们构建了一种 HSV 载体,其 EPO 的表达受 Tet-on 系统的控制,其中 HSV 潜伏相关启动子 2 元件用于驱动 Tet-on 转录激活剂的表达。该载体的 EPO 表达可通过给予强力霉素(DOX)进行体外严密调控。载体接种后 1 个月,可在体内转导背根神经节(DRG),给予含 DOX 的饲料可诱导 EPO 的表达。连续给予含 DOX 的饲料 3 个月,可保护链脲佐菌素诱导糖尿病小鼠免受神经病变的发生。在糖尿病的 3 个月中,每两周给予 DOX 也可达到相同的效果,但每周给予 DOX1 次仅能部分保护神经病变的发生。综上,这些结果表明,这种载体非常适合临床试验,用于治疗慢性或亚急性进展性神经病变。